Protein kinases are utilized by cells during cell division and development, cytokinesis, and for the maintenance of metabolic homeostasis. Given the well-established role of Aurora kinase B (AurkB) in cellular division and cancer development, we have made a surprising recent discovery – that AurkB has a critical role in axonal outgrowth of neurons (a non-dividing cell). We have recently identified AurkB in neuronal regeneration [1] but its substrates involved in neurite outgrowth remains to be investigated.  Here, we aim to explore the potential phosphorylated substrates of AurkB and their involvement in neuritogenesis. Utilizing titanium dioxide enrichment and LC-MS/MS, differentiated RGC-5 cells grown in SILAC neurobasal media were treated with AurkB inhibitor, AZD1152-HQPA, and cell lysates were digested with trypsin and enriched for phosphopeptides. We identified >10,000 phosphopeptides, in which ~650 phosphopeptides were differentially down-regulated upon treatment. Preliminary analysis revealed that some key proteins involved in axonal transport are under the regulation of AurkB. The role of AurkB was further supported by in vivo studies of transgenic zebrafish, demonstrating that overexpression of AurkB regulates axonal elongation and neurite outgrowth. Collectively, this data suggests that apart from cellular division, AurkB could play a critical role in the development of neurons.